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Cloning and Characterization a Prokaryotic Pantothenate Kinase Gene (panK ) from Streptomyces peuceticus ATCC27952

초록

영어

Coenzyme A (CoA) is a ubiquitous essential cofactor. CoA is the source of 4` -phosphopantetheine, the prosthetic group of carrier proteins of fatty acid, polyketide and nonribosomal peptide synthases(1). Pantothenate kinase (PanK) is the key regulatory enzyme in the CoA biosynthetic pathway(2). Most polyketides are synthesized by the assembly of precursors such as acethyl-, propionyl- and butyl-CoA. Their polyketide synthesis precursors are activated derivatives, malonyl-, methylmalnoly- and ethylmalonyl-CoA. Recently, the pantothenate kinase gene cluster was analyzed from S. peucetius genome project. The
multiple amino acid sequence of SpPanK shows 52% and 58% similarity with the comparison of EcPanK and M tPanK. We cloned panK gene from S. peucetius ATCC 27952 and constructed the expression vector in pET32a. The expressed protein is 36 kDa. The enzyme assay was monitored to ADP formation by coupling reactions with pyruvate kinase and lactate dehydrogenase. The activity was measured as a change in absorbance at 340 nm during
monitoring of depletion of NADH(3).

저자정보

  • Eung Pil Kim Institute of Biomolecule Reconstruction (iBR), Department of Biological-Engineering, Sunmoon University
  • Hei Chan Lee Institute of Biomolecule Reconstruction (iBR), Department of Biological-Engineering, Sunmoon University
  • Tae-Jin Oh Institute of Biomolecule Reconstruction (iBR), Department of Biological-Engineering, Sunmoon University
  • Jae Kyung Sohng Institute of Biomolecule Reconstruction (iBR), Department of Biological-Engineering, Sunmoon University

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